Symphony ANA Screening

Symphony ANA Screening

Surer Screening for Connective Tissue Diseases High clinical relevance high sensitivity supports diagnosis high specificity avoids inappropriate follow-up High technical performance low variances and high reproducibility for consistent results high lot-to-lot consistency due to validated production procedures semi-quantitative results expressed as a ratio relative to a defined calibrator State-of-the-art antigens human recombinant U1RNP (mixture of recombinant RNP70, A, C) highly purified SmD protein human recombinant Ro (52, 60kDa) human recombinant La human recombinant Scl-70 human recombinant CENP-B human recombinant Jo-1 Automation screen and individual specificity testing available in one run choice of ImmunoCAP TM instruments available for low to high throughput stored standard curve to be used with all EliA TM IgG analytes urgent samples can be run cost-efficiently reflex testing from screen to single-specificity assays Easy handling serum as well as plasma can be used automated sample dilution

Picture 1: SLE, pleural effusions in lung x-ray Connective Tissue Diseases Connective Tissue diseases (CTDs) represent classical models of systemic autoimmune diseases. They are a heterogeneous group of diseases characterised by abnormal structure or function of one or more of the elements of connective tissue, i.e. collagen, elastin or the mucopolysaccharides. Differential diagnosis of CTDs is mainly based on clinical findings but is complicated by the similarity of their symptoms. Therefore, autoantibodies are useful markers to support the diagnosis or exclusion of CTDs. The most prominent CTDs are systemic lupus erythematosus (SLE; potentially affecting all organs), Sjögren s syndrome (SS; characterised by diminished lacrimal and salivary gland secretions), scleroderma (systemic sclerosis, SSc; a chronic, progressive dermatosis), limited systemic sclerosis (a scleroderma formerly known as CREST syndrome, with a more benign disease course), poly-/dermatomyositis (PM/DM; an acute or chronic inflammatory disease of muscle and skin), and mixed connective tissue disease (MCTD; a syndrome with features of scleroderma, rheumatoid arthritis, SLE and PM/DM). Picture 2: Picture 3: Raynaud s phenomenon Dermatomyositis, calcification Disease Markers The presence and specificity of certain autoantibodies give a strong indication as to the likely CTD involved. The prevalence of marker autoantibodies in particular CTDs are summarised in Table 1. Marker Autoantibody Associated CTD Autoantibody Prevalence U1RNP MCTD, SLE 100%, 30 70% Sm SLE 20 30% SS-A/Ro Sjögren s syndrome, SLE 60 90%, 25 30% SS-B/La Sjögren s syndrome, SLE 40 95%, 6 15% Scl-70 Systemic sclerosis 20 70% CENP Limited systemic sclerosis (CREST) 70 80% Jo-1 Poly-/dermatomyositis 25 35% Table 1: Prevalence of autoantibodies in connective tissue diseases. Antigens Recombinant antigens from a eukaryotic system for best quality, consistency and performance was developed clinically in order to maximise the assay s usefulness in a diagnostic setting. All antigens included have been selected on the basis of the related antibody s significance in one or more of the connective tissue diseases. The result is a clinically relevant, sensitive and highly specific screening assay. As an intact three-dimensional structure of the antigens (conformation) is crucial for recognition by antibodies, our human recombinant antigens are produced in the eukaryotic baculovirus/insect cells system which, in contrast to bacterial systems, is capable of expressing the antigens in the correct conformation and performing the complex posttranslational modifications necessary to ensure the protein is antigenically identical to the human native form. Using recombinant antigens wherever possible allows us to minimise contaminants, avoid harsh, protein-altering purification processes and guarantee a high lot-to-lot consistency of the antigens.

Sensitive screening with high specificity High Clinical Relevance Pos Neg SLE 21 13 (incl 7 patients with inactive disease) Other CTD 13 7 Other AI Disease 2 24 Non AI Disease 8 82 Totals 44 126 Table 2: Clinical performance of EliA TM Symphony (Gonzalez et al 2005; Clin Chim Acta 359:109-114) The results described in this paper demonstrate that has an extremely good clinical performance with a PPV of 77% and NPV of 84%. The Positive Likelihood Ratio of 7.3 for EliA Symphony compared favourably with that of the less specific HEp-2 IIF at 6.5 (IIF cut-off at 1:160). Targeted antigen selection for maximum clinical relevance In a further study, using samples previously determined to contain specific ENA antibodies, EliA Symphony showed outstanding sensitivity in detecting the antibodies. Of particular note are the results for SS-A/Ro and Jo-1 antibodies which are typically difficult to detect using IIF methods. No. of Samples Target Specificity No. Correctly Identified by 51 SS-A/Ro 51 45 SS-B/La 45 44 RNP 44 5 Sm 5 17 Jo-1 17 13 Scl-70 12 Table 3: Performance of in 175 sera with predefined specificities (Oris et al 2002; Poster presented at the 6th Dresden Symposium on Autoantibodies) IgG levels calibrated against International Reference Preparation (IRP) 67/86 from World Health Organization CDC Serum Target Result (Neg <0.7 Pos >1.0) CDC1 Homogeneous/Rim 1.1 CDC2 Speckled La 21.8 CDC3 Speckled 37.6 CDC4 U1-RNP 12.9 CDC5 Sm 12.0 CDC6 Nucleolar 0.2 CDC7 SS-A/Ro 13.4 CDC8 Centromere 8.6 CDC9 Scl-70 5.9 CDC10 Jo-1 16.4 AMLI Panel Member Target Result (Neg <0.7 Pos >1.0) A CENP 2.7 B Scl-70 4.6 D RNP 6.6 E SS-A/Ro 2.0 F Jo-1 4.3 G SS-B/La 31.3 I Sm 30.9 J dsdna 2.7 K Neg 0.0 L Neg 0.0 Table 4: Performance of with reference preparations All CDC sera are found positive with the exception of CDC6. This serum contains antibody specificities directed to antigens which are not included in the test. finds all AMLI panel sera correctly positive or negative as defined in the targets. The positive response for Member J is due to the documented presence of Ro and RNP antibodies in this sample as antibodies to dsdna are not detected by this system.

The EliA System Time for the Essentials completely automated (true walk-away, overnight runs) easy instrument management by custom-made software barcode-reader (optional for ImmunoCAP TM 100 E ) protocols, QC and raw data easily accessible optional host link detailed QC management integrated stock management system on the ImmunoCAP TM 250 Cost efficient and flexible autoimmunity and allergy on the same instrument different autoimmune tests in the same run (puzzle-kit-approach) no batching of samples necessary small runs can be handled cost-effectively once-monthly calibration curve control each run several ImmunoCAP TM instruments can be linked A boost in service for your laboratory and your clinicians sample result turnaround the same day STAT function on ImmunoCAP TM 250 for immediate testing of emergency samples overnight runs possible detailed documentation of results (patient or requester specific) ImmunoCAP TM 100 E up to 46 determinations in less than 2.5 hours ImmunoCAP TM 250 fully automated random access up to 350 determinations per shift multiple methods in one run positive identification and traceability of samples and reagents on ImmunoCAP TM 250

Technical Data Product Antigens Cut-off Measuring Range Dilution Sample Material Normal Distribution human recombinant U1RNP (70kDa, A, C), Ro (60kDa, 52kDa), La, Centromere B, Scl-70 and Jo-1 proteins, native purified Sm proteins neg. < 0.7; equiv. 0.7 1.0; pos>1.0 (Ratio) 0.03 (at least) 32 (Ratio) 1:100 (automated) Serum, Plasma (EDTA, citrate, heparin) Mean 0.2, 95 th percentile 0.4 (Ratio) Reproducibility Intra-run CV* 3.7 8.8 % Inter-run CV* 0.0 4.8 % *for details see Directions For Use Ordering Information Package size Article No. Well 4 x 12 14-5508-01 EliA ANA Control (neg + pos) 2 x 3 83-1004-01 EliA IgG Calibrator Well 4 x 12 14-5509-01 EliA 100 Reagents EliA IgG Conjugate 6 x 48 83-1002-01 EliA IgG Conjugate 2 x 48 83-1005-01 EliA IgG Calibrator 6 vials 83-1000-01 EliA IgG Curve Control 6 vials 83-1001-01 EliA 250 Reagents EliA IgG Conjugate 50 6 x 50 83-1017-01 EliA IgG Conjugate 200 6 x 200 83-1018-01 EliA IgG Calibrator Strips 5 strips 83-1015-01 EliA IgG Curve Control Strips 5 strips 83-1016-01 General Reagents ImmunoCAP 100 ImmunoCAP Development Kit 10-9263-01 ImmunoCAP Washing Solution 6 x 1 l 10-9422-01 General Reagents ImmunoCAP 250 ImmunoCAP Development Solution 6 x 180 10-9440-01 ImmunoCAP Stop Solution 6 x 590 10-9442-01 ImmunoCAP Washing Solution 2 x 5 l 10-9202-01 Phadia GmbH, Munzinger Str. 7, D-79111 Freiburg Germany, Tel: +49 761 47 805 0, Fax: +49 761 47 805 338, autoimmunity@phadia.com, www.phadia.com Head office Sweden +46 18 16 50 00 Austria +43 1 270 20 20 Belgium +32 2 749 55 15 Brazil +55 11 3345 5050 Denmark +45 70 23 33 06 Finland +358 9 8520 2560 France +33 1 6137 3430 Germany +49 761 47 805 0 Italy +39 02 64 163 411 Japan +81 3 5365 8332 Norway +47 21 67 32 80 Portugal +351 21 423 53 50 Spain +34 935 765 800 Distributors +46 18 16 50 00 Switzerland +41 43 343 40 50 Taiwan +886 2 2516 0925 The Netherlands +31 30 602 37 00 United Kingdom / Ireland +44 1908 84 70 34 USA +1 800 346 4364 Other Countries +46 18 16 30 00 Order No. 91 51 508 Freiburg 08/2006